| 人ski基因真核表达载体的构建及促细胞增殖作用 |
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| 引用本文： | 彭艳,李平,刘苹,陈磊,赵晓光,周元国. 人ski基因真核表达载体的构建及促细胞增殖作用[J]. 中国组织工程研究与临床康复, 2011, 15(41): 7693-7696. DOI: 10.3969/j.issn.1673-8225.2011.41.023 |
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| 作者姓名： | 彭艳 李平 刘苹 陈磊 赵晓光 周元国 |
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| 作者单位： | 解放军第三军医大学附属大坪医院野战外科研究所分子生物学中心,创伤、烧伤与复合伤国家重点实验室,重庆市400042 |
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| 基金项目： | 创伤、烧伤与复合伤国家重点实验室自主课题 |
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| 摘 要： | 背景:c-Ski既可促进组织修复,又能降低瘢痕形成,有望成为一个全新的基因治疗药物。目的:构建人ski基因的真核表达载体,并对其促成纤维细胞增殖效果进行验证。方法:RT-PCR法从人包皮培养原代成纤维细胞总RNA中扩增出人ski基因,连同真核表达启动子CMV克隆到pUC118载体上,酶切和测序验证后,应用脂质体将其转染到培养的大鼠皮肤成纤维细胞中。结果与结论:酶切和测序结果证实表达质粒构建成功,Westernblot显示Ski蛋白在转染细胞中成功表达,且可显著提高转染细胞的增殖效应,说明以pUC118为骨架的重组ski表达质粒具有促进大鼠皮肤成纤维细胞增殖的作用。
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| 关 键 词： | 载体构建 ski pUC118 成纤维细胞 增殖 |
Construction of eukaryotic expression vecter for human ski gene and observation of its role in promoting cell proliferation |
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| Peng Yan,Li Ping,Liu Ping,Chen Lei,Zhao Xiao-guang,Zhou Yuan-guo. Construction of eukaryotic expression vecter for human ski gene and observation of its role in promoting cell proliferation[J]. Journal of Clinical Rehabilitative Tissue Engineering Research, 2011, 15(41): 7693-7696. DOI: 10.3969/j.issn.1673-8225.2011.41.023 |
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| Authors: | Peng Yan Li Ping Liu Ping Chen Lei Zhao Xiao-guang Zhou Yuan-guo |
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| Affiliation: | Peng Yan,Li Ping,Liu Ping,Chen Lei,Zhao Xiao-guang,Zhou Yuan-guo State Key Laboratory of Trauma,Burns and Combined Injury,Center for Molecular Biology,Research Institute of Surgery and Daping Hospital,Third Military Medical University of Chinese PLA,Chongqing 400042,China |
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| Abstract: | BACKGROUND:c-Ski can promote tissue repair and alleviate scar formation,which is expected to be a new gene therapeutic drugs.OBJECTIVE:To construct eukaryotic expression plasmid of human ski gene and to investigate its proliferative role in fibroblasts.METHODS:The ski gene was amplified by polymerase chain reaction(PCR) from human foreskin cultured fibroblast and was sub-cloned into pUC118 with CMV eukaryotic promoter.The recombinant pUC118-Ski plasmids were transfected into rat skin fibroblasts using lipof... |
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